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Performance evaluation and methods comparison of transcriptomic-based approaches for the characterization of wastewater treatment effluent

Wastewater treatment effluents (WWTE) present complex risks to aquatic ecosystems that are difficult to characterize using traditional methods. This study systematically evaluated the consistency and performance of transcriptomic-based approaches over time with repeated sampling and with differing experimental approaches (selection of reference condition, grab vs. composite sampling, deployed vs. laboratory exposed). RNA-seq was performed on larval fathead minnow (FHM) exposed in the laboratory to moderately hard reconstituted water (MRHW) or individual grab samples collected from an upstream site and a WWTE in the morning and afternoon over two successive days, as well as FHM deployed concurrently with grab sampling at the same sites. Composite transcriptional profiles were generated by pooling count data from grab sample exposures. The choice of comparator significantly affected results. The use of the upstream site as the reference consistently yielded fewer differentially expressed genes (DEGs) and minimal overlap compared to DEG sets from the other comparisons. Using MRHW as a comparator, DEG sets showed high consistency across grab samples, with morning samples demonstrating larger, highly consistent gene expression sets (96 % overlap) compared to afternoon samples, revealing clear and consistent within-day expression patterns. With the MHRW comparator, DEG sets from grab sample composites and deployments also overlapped substantially, indicating that transcriptional profiles accurately reflect WWTE composition regardless of exposure method. Comparisons with non-targeted (NTA) and targeted analytical datasets confirmed that gene expression interpretations aligned with effluent composition while highlighting limitations of relying solely on targeted analyte sets for connecting expression to specific chemicals. Though highly dependent on experimental design, these results demonstrate that transcriptomic-based methods provide significant utility for characterizing the bioactivity of complex environmental mixtures, and when paired with NTA datasets, have the potential to deliver a comprehensive assessment of their environmental risk.

Environmental Pollution↗

Selenium and mercury tissue partitioning and trophodynamics in the Lake Koocanusa (USA–Canada) fish community

Mining-related contaminants such as selenium (Se) and mercury (Hg) accumulate in aquatic organisms and transfer through aquatic food webs, where they can exert toxic effect undermining the ecological health of aquatic resources. Yet, how Se and Hg co-distribute within food webs and within individual organisms remains poorly understood. We compiled muscle and ovary Se and Hg concentration data from fishes captured in Lake Koocanusa—a North American reservoir receiving elevated Se and Hg inputs from coal mining operations in the Elk River Valley, Canada—to provide insights into the co-processing of Se and Hg within fish and among fish species, including how ecological or seasonal factors shift these distribution patterns. Se and Hg concentrations (ranges: 0.21–38.7 μg Se g −1 dry weight and 0.01–0.84 μg Hg g −1 wet weight), as well as Se:Hg molar ratios, varied substantially between tissues and among species, reflecting biological and ecological differences. Our results suggest that the decoupling of Se or Hg deposition in fish muscle versus ovary may reflect different pathways of impairment, at the individual and population levels, and that interspecific and tissue-specific variability makes it challenging to set a universal Se:Hg threshold for the protection of aquatic organisms.

British Columbia, Montana↗

Per- and polyfluoroalkyl substances and pesticides in black bears (Ursus americanus) and grizzly bears (Ursus arctos horribilis) from Montana's Cabinet-Yaak ecosystem: A baseline assessment of emerging and legacy contaminants

Per- and polyfluoroalkyl substances (PFAS) and pesticides are globally distributed contaminants that persist in terrestrial food webs, yet baseline data for large omnivores in the continental U.S. remain limited. This study quantified PFAS and pesticides in whole blood from free-ranging black bears ( Ursus americanus , n = 15) and grizzly bears ( Ursus arctos horribilis , n = 9) in the Cabinet-Yaak Ecosystem of northwestern Montana. Using high-resolution mass spectrometry-based methods (GC-MS/MS and LC-MS/MS), 52 PFAS and 186 pesticides were analyzed. PFAS were detected in all bears, with total PFAS concentrations ranging from 79.5 to 317 pg/mL. PFAS profiles were dominated by long-chain perfluoroalkyl carboxylic acids, particularly PFOA, PFDA, and PFUnDA, with minimal contribution from short-chain compounds. Species was the strongest ecological predictor of PFAS concentrations, although variability across age, sex, and capture location were observed. Only two pesticides, piperonyl butoxide and permethrin, were detected at quantifiable concentrations (2070–12,600 pg/mL), and no correlations were observed between pesticide and PFAS concentrations, indicating independent exposure pathways. The predominance of long-chain PFAS suggests diffuse environmental sources and bioaccumulation within terrestrial food webs. Although measured concentrations were generally low relative to other wildlife toxicological studies, the persistence and protein-binding properties of long-chain PFAS warrant continued monitoring, particularly for long-lived omnivores. These results establish baseline contaminant concentrations for two apex omnivores in a remote ecosystem and highlight the utility of large mammals as integrative sentinels of environmental contaminant exposure for regional monitoring and wildlife conservation. Continued monitoring in the Cabinet-Yaak and across western North America will be critical for assessing temporal trends, identifying new contaminant sources, and evaluating ecological health in bear populations.

Montana↗

Use of radium isotopes to determine the age and origin of radioactive barite at oil-field production sites

Radium-bearing barite (radiobarite) is a common constituent of scale and sludge deposits that form in oil-field production equipment. The barite forms as a precipitate from radium-bearing, saline formation water that is pumped to the surface along with oil. Radioactivity levels in some oil-field equipment and in soils contaminated by scale and sludge can be sufficiently high to pose a potential health threat. Accurate determinations of radium isotopes (226Ra+228Ra) in soils are required to establish the level of soil contamination and the volume of soil that may exceed regulatory limits for total radium content. In this study the radium isotopic data are used to provide estimates of the age of formation of the radiobarite contaminant. Age estimates require that highly insoluble radiobarite approximates a chemically closed system from the time of its formation. Age estimates are based on the decay of short-lived 228Ra (half-life=5.76 years) compared to 226Ra (half-life=1600 years). Present activity ratios of 228Ra/226Ra in radiobarite-rich scale or highly contaminated soil are compared to initial ratios at the time of radiobarite precipitation. Initial ratios are estimated by measurements of saline water or recent barite precipitates at the site or by considering a range of probable initial ratios based on reported values in modern oil-field brines. At sites that contain two distinct radiobarite sources of different age, the soils containing mixtures of sources can be identified, and mixing proportions quantified using radium concentration and isotopic data. These uses of radium isotope data provide more description of contamination history and can possibly address liability issues. Copyright ?? 2000 .

Environmental Pollution↗

Use of passive ambient ozone (O3) samplers in vegetation effects assessment

A stochastistic, Weibull probability model was developed and verified to simulate the underlying frequency distributions of hourly ozone (O3) concentrations (exposure dynamics) using the single, weekly mean values obtained from a passive (sodium nitrite absorbent) sampler. The simulation was based on the data derived from a co-located continuous monitor. Although at the moment the model output may be considered as being specific to the elevation and location of the study site, the results were extremely good. This effort for the approximation of the O3 exposure dynamics can be extended to other sites with similar data sets and in developing a generalized understanding of the stochastic O3 exposure-plant response relationships, conferring measurable benefits to the future use of passive O3 samplers, in the absence of continuous monitoring. Copyright ?? 2000 Elsevier Science Ltd.

Environmental Pollution↗

Environmental contaminants in prey and tissues of the peregrine falcon in the Big Bend Region, Texas, USA.

Peregrine falcons ( Falco peregrinus ) have been recorded nesting in Big Bend National Park, Texas, USA and other areas of the Chihuahuan Desert since the early 1900s. From 1993 to 1996, peregrine falcon productivity rates were very low and coincided with periods of low rainfall. However, low productivity also was suspected to be caused by environmental contaminants. To evaluate potential impacts of contaminants on peregrine falcon populations, likely avian and bat prey species were collected during 1994 and 1997 breeding seasons in selected regions of western Texas, primarily in Big Bend National Park. Tissues of three peregrine falcons found injured or dead and feathers of one live fledgling also were analyzed. Overall, mean concentrations of DDE [1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene], a metabolite of DDT [1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane], were low in all prey species except for northern rough-winged swallows ( Stelgidopteryx serripennis , mean = 5.1 microg/g ww). Concentrations of mercury and selenium were elevated in some species, up to 2.5 microg/g dw, and 15 microg/g dw, respectively, which upon consumption could seriously affect reproduction of top predators. DDE levels near 5 microg/g ww were detected in carcass of one peregrine falcon found dead but the cause of death was unknown. Mercury, selenium, and DDE to some extent, may be contributing to low reproductive rates of peregrine falcons in the Big Bend region.

Texas↗

Productivity responses of Acer rubrum and Taxodium distichum seedlings to elevated CO2 and flooding

Elevated levels of atmospheric CO2 are expected to increase photosynthetic rates of C3 tree species, but it is uncertain whether this will result in an increase in wetland seedling productivity. Separate short-term experiments (12 and 17 weeks) were performed on two wetland tree species, Taxodium distichum and Acer rubrum, to determine if elevated CO2 would influence the biomass responses of seedlings to flooding. T. distichum were grown in replicate glasshouses (n = 2) at CO2 concentrations of 350 or 700 ppm, and A. rubrum were grown in growth chambers at CO2 concentrations of 422 or 722 ppm. Both species were grown from seed. The elevated CO2 treatment was crossed with two water table treatments, flooded and non-flooded. Elevated CO2 increased leaf-level photosynthesis, whole-plant photosynthesis, and trunk diameter of T. distichum in both flooding treatments, but did not increase biomass of T. distichum or A. rubrum. Flooding severely reduced biomass, height, and leaf area of both T. distichum and A. rubrum. Our results suggest that the absence of a CO2-induced increase in growth may have been due to an O2 limitation on root production even though there was a relatively deep (??? 10 cm) aerobic soil surface in the non-flooded treatment. ?? 2001 Elsevier Science Ltd. All rights reserved.

Environmental Pollution↗

Dioxins and congener-specific polychlorinated biphenyls in three avian species from the Wisconsin River, Wisconsin

Sediments from the Wisconsin River, WI, USA are contaminated with 2,3,7,8-tetrachloro- p -dioxin (TCDD) and polychlorinated biphenyls (PCBs). Wet weight concentrations of TCDD and PCBs in eggs were at background levels and highest in the piscivorous hooded merganser ( Lophodytes cucullatus ; geometric mean=7 pg/g TCDD and 0.92 μg/g PCBs) and lowest in the omnivorous wood duck ( Aix sponsa ) (<1 pg/g and 0.07 μg/g); concentrations in eggs of the insectivorous tree swallow ( Tachycineta bicolor ) were intermediate (<1 pg/g and 0.33 μg/g). Positive accumulation rates of TCDD (8–19 pg/day) and PCBs (0.4–0.7 μg/day) in tree swallow nestlings suggest that the Wisconsin River is the source of these contaminants for tree swallow nestlings. The lower representation of trichlorobiphenyls and tetrachorobiphenyls in hooded merganser eggs compared to wood duck or tree swallow eggs suggests that the hooded merganser or its diet has a greater ability to metabolize lower-numbered PCB congeners than wood ducks or tree swallows.

Wisconsin↗

Arsenic species and chemistry in groundwater of southeast Michigan

Groundwater samples, taken from 73 wells in 10 counties of southeast Michigan in 1997 had arsenic concentrations in the range of 0.5 to 278 μg/l, the average being 29 μg/l. About 12% of these wells had arsenic concentrations that exceeded the current USEPA's maximum contaminant level of 50 μg/l. Most (53-98%) of the arsenic detected was arsenite [As(III)] and other observations supported the arsenic species distribution (low redox potential and DO). In shallow groundwater (15 m), the concentration of arsenic is possibly controlled by reductive dissolution of arsenic-rich iron hydroxide/oxyhydroxide and dissolution of arsenic sulfide minerals.

Environmental Pollution↗

Impacts of swine manure pits on groundwater quality

Manure deep-pits are commonly used to store manure at confined animal feeding operations. However, previous to this study little information had been collected on the impacts of deep-pits on groundwater quality to provide science-based guidance in formulating regulations and waste management strategies that address risks to human health and the environment. Groundwater quality has been monitored since January 1999 at two hog finishing facilities in Illinois that use deep-pit systems for manure storage. Groundwater samples were collected on a monthly basis and analyzed for inorganic and bacteriological constituent concentrations. The two sites are located in areas with geologic environments representing different vulnerabilities for local groundwater contamination. One site is underlain by more than 6 m of clayey silt, and 7-36 m of shale. Concentrations of chloride, ammonium, phosphate, and potassium indicated that local groundwater quality had not been significantly impacted by pit leakage from this facility. Nitrate concentrations were elevated near the pit, often exceeding the 10 mg N/l drinking water standard. Isotopic nitrate signatures suggested that the nitrate was likely derived from soil organic matter and fertilizer applied to adjacent crop fields. At the other site, sandstone is located 4.6-6.1 m below land surface. Chloride concentrations and ??15N and ??18O values of dissolved nitrate indicated that this facility may have limited and localized impacts on groundwater. Other constituents, including ammonia, potassium, phosphate, and sodium were generally at or less than background concentrations. Trace- and heavy-metal concentrations in groundwater samples collected from both facilities were at concentrations less than drinking water standards. The concentration of inorganic constituents in the groundwater would not likely impact human health. Fecal streptococcus bacteria were detected at least once in groundwater from all monitoring wells at both sites. Fecal streptococcus was more common and at greater concentrations than fecal coliform. The microbiological data suggest that filtration of bacteria by soils may not be as effective as commonly assumed. The presence of fecal bacteria in the shallow groundwater may pose a significant threat to human health if the ground water is used for drinking. Both facilities are less than 4 years old and the short-term impacts of these manure storage facilities on groundwater quality have been limited. Continued monitoring of these facilities will determine if they have a long-term impact on groundwater resources. ?? 2002 Elsevier Science Ltd. All rights reserved.

Environmental Pollution↗

Effects of salinity, pH and temperature on the re-establishment of bioluminescence and copper or SDS toxicity in the marine dinoflagellate Pyrocystis lunula using bioluminescence as an endpoint

Pyrocystis lunula is a unicellular, marine, photoautotrophic, bioluminescent dinoflagellate. This organism is used in the Lumitox ?? bioassay with inhibition of bioluminescence re-establishment as the endpoint. Experiments determined if acute changes in pH, salinity, or temperature had an effect on the organisms' ability to re-establish bioluminescence, or on the bioassay's potential to detect sodium dodecyl sulfate (SDS) and copper toxicity. The re-establishment of bioluminescence itself was not very sensitive to changes in pH within the pH 6-10 range, though reducing pH from 8 to levels below 6 decreased this capacity. Increasing the pH had little effect on Cu or SDS toxicity, but decreasing the pH below 7 virtually eliminated the toxicity of either compound in the bioassay. Lowering the salinity from 33 to 27??? or less resulted in a substantial decrease in re-establishment of bioluminescence, while increasing the salinity to 43 or 48 ??? resulted in a small decline. Salinity had little influence on the bioassay's quantification of Cu toxicity, while the data showed a weak negative relationship between SDS toxicity and salinity. Re-establishment of bioluminescence showed a direct dependence on temperature, but only at 10??C did temperature have an obvious effect on the toxicity of Cu in this bioassay. ?? 2003 Elsevier Science Ltd. All rights reserved.

Environmental Pollution↗

Groundwater quality surrounding Lake Texoma during short-term drought conditions

Water quality data from 55 monitoring wells during drought conditions surrounding Lake Texoma, located on the border of Oklahoma and Texas, was compared to assess the influence of drought on groundwater quality. During the drought month of October, water table levels were three feet (0.9 m) lower compared with several months earlier under predrought climate conditions. Detection frequencies of nitrate (> 0.1 mg/l), orthophosphates (> 0.1 mg/l), chlorides (> MCL), and sulfates (> MCL) all increased during drought. Orthophosphate level was higher during drought. Largest increases in concentration were nitrate under both agriculture lands and in septic tank areas. An increase in ammonium-nitrogen was only detected in the septic tank area. The study showed that stressors such as nitrate and total salts could potentially become a health or environmental problem during drought.

Environmental Pollution↗

Effects of the mosquito larvicide GB-1111 on red-winged blackbird embryos

Golden Bear Oil ( GB - 1111 ; legal trade name for GB -1313) is a petroleum distillate that is used in the United States and other countries as a larvicide for mosquito suppression. As part of a multi-species evaluation of the potential effects of GB - 1111 on birds, red - winged blackbird eggs were collected, artificially incubated, and treated with one of five amounts of GB - 1111 varying from 0 to 10 times the expected exposure from a spray application of the maximum recommended amount (X=47 l/ha, 5 gal/ac). The application of 10 X caused a significant reduction in hatching success. A dose-related reduction of hepatic microsomal monooxygenase activity (EROD) was detected. Among body weights, skeletal measurements, and age at death, only crownrump length was different among experimental groups. Overall, the potential hazard to embryos of a representative wetland passerine appears minimal until the application rate exceeds 3 X.

Environmental Pollution↗

Heavy metals and metalloids in egg contents and eggshells of passerine birds from Arizona

Concentrations of inorganic elements were determined in eggs of passerine birds including the endangered southwestern willow flycatcher ( Empidonax traillii extimus ) from four regions in Arizona. The main aim of the study was to determine the distribution of metals in egg contents and eggshells, with emphasis on the deposition of Sr in eggshells. Seventy eggs of 11 passerine species were collected at four nesting locations during 2000. Aluminum, Ba, Cr, Cu, Mn, Se, Sr, and Zn, were detected primarily in egg contents of all bird species. Arsenic, Ni, Pb, and V were detected primarily in eggshells. A proportion of most inorganic elements accumulated in the eggshell. Concentrations of Ba, Cu, Mn, Se, Sr, and Zn in egg contents and As, Ba, Cu, and V in eggshells of yellow-breasted chats ( Icteria virens ) were similar among locations. However, concentrations of Mn, Ni, Sr, and Zn in eggshells were significant different among locations. Except for Cu, Mn, Se, and Zn, concentrations of inorganic elements were 2–35 times greater in eggshells than in eggs. Most concentrations of metals and metalloids in eggs and eggshells of all the bird species were below levels known to affect reproduction or that have other deleterious effects. However, I found somewhat elevated concentrations of Sr in eggshells (highest mean=1505 μg/g dw, n =3) of yellow-breasted chats and willow flycatchers, and in egg contents of yellow warblers ( Dendroica petechia) and song sparrows ( Melospiza melodia ). Whether current observed concentrations of Sr in eggshells are affecting nesting birds in Arizona remains to be determined. Strontium and other metals could be associated with lower hatching success in some areas. This study shows that a proportion of many inorganic elements accumulates in the eggshell and that the potential effects on the proper structure and functioning of the eggshell should not be ignored.

Environmental Pollution↗

Effects of lead-contaminated sediment and nutrition on mallard duckling brain growth and biochemistry

Day-old mallard ( Anas platyryhnchos ) ducklings received either a clean sediment (24%) supplemented control diet, Coeur d'Alene River Basin, Idaho (CDARB) sediment ( 3449 μg/g lead ) supplemented diets at 12% or 24%, or a positive control diet (24% clean sediment with equivalent lead acetate to the 24% CDARB diet) for 6 weeks. The 12% CDARB diet resulted in a geometric mean concentration of 396 ppb (WW) brain lead with decreased brain protein and ATP concentrations but increased oxidized glutathione (GSSG) relative to the control diet. The 24% CDARB diet resulted in a concentration of 485 ppb brain lead with lower brain weight and ATP concentration than controls but higher concentrations of reduced glutathione (GSH) and calcium. Lead acetate accumulated twice as well as CDARB derived lead and resulted in histopathological lesions of the brain. With a combination of a suboptimal diet and 24% CDARB, brain lead concentration was higher (594 ppb) than with 24% CDARB in the standard diet, histopathological lesions became apparent and GSH was higher than suboptimal diet controls.

Idaho↗

Clarification of effects of DDE on shell thickness, size, mass, and shape of avian eggs

Moriarty et al. (1986) used field data to conclude that DDE decreased the size or altered the shape of avian eggs; therefore, they postulated that decreased eggshell thickness was a secondary effect because, as a general rule, thickness and egg size are positively correlated. To further test this relationship, the present authors analyzed data from eggs of captive American kestrels. Falco sparverius given DDT- or DDE-contaminated or clean diets and from wild brown pelicans Pelecanus occidentalis collected both before (pre-1946) and after (post-1945) DDT was introduced into the environment. Pertinent data from other field and laboratory studies were also summarized. DDE was not related to and did not affect size, mass, or shape of eggs of the brown pelican or American kestrel; but the relationship of DDE to eggshell thinning held true. Size and shape of eggs of brown pelicans from the post-1945 era and those of kestrels, on DDT-contaminated diets showed some significant, but inconsistent, changes compared to brown pelican data from the pre-1946 era or kestrels on clean diets. In contrast, nearly all samples of eggs of experimental kestrels given DDT-contaminated diets and those of wild brown pelicans from the post-1945 era exhibited significant eggshell thinning. Pertinent experimental studies with other sensitive avian species indicated no effects of DDE on the size or shape of eggs, even though the high dietary concentrations caused extreme eggshell thinning and mortality of some adult mallards (Anas platyrhynchos) in one study. These findings essentially controvert the argument that decreased eggshell thickness is a secondary effect resulting from the primary effect of DDE-induced changes in the size or shape of eggs.

Environmental Pollution↗

Spatial and temporal heterogeneity in metallic elements in industrialized aquatic bird habitat

This study examined the potential for exposure of migratory aquatic birds to contaminants in highly industrialized habitats at Baltimore Harbor, Maryland. Seven nearshore, benthic sites were sampled every 3 or 6 months from June 1987 to December 1988. Twenty metallic elements were measured in composites (i.e. annelids, amphipods, and isopods were pooled), clams, and sediments. Elevated concentrations were widespread throughout the sites, depending on the element. Most concentrations in composites were lowest at sites innermost and outside the harbor. Higher concentrations in composites were not related to season. Most concentrations were higher in composites than in sediments or clams, but they varied by element for clams and sediments. The largest flock of wintering waterfowl frequented the site exhibiting the highest concentrations of most elements ( p < 0.0001). Average concentrations of known toxic elements were probably not harmful to aquatic birds or their prey, but the highest detected concentrations of eight elements warrant caution. At least eight other elements exceeded background concentrations, but toxicity to birds or their prey are unknown.

Maryland↗

Hydrophobic organochlorine compounds sequestered in submersed aquatic macrophytes (Hydrilla yerticillata (L.f.) Royle) from the tidal Potomac River (USA)

The potential for hydrophobic organochlorine contaminants to be sequestered in submersed aquatic vegetation was evaluated by determining the concentrations of cis - and trans -chlordane, dieldrin, and polychlorinated biphenyls (PCBs) in feral aquatic macrophytes ( Hydrilla verticillata (L.f.) Royle) collected from the tidal Potomac River. Similarities in mean dry-weight concentrations of the identified organochlorine compounds in H. verticillata and surrounding alluvial sediments indicated that the extent of sequestration in H. verticillata was of the same magnitude as sorption of these compounds to river sediments, but some qualitative differences in PCB congener profiles existed. The results imply that to some degree H. verticillata can influence downstream fluxes of organic contaminants in fluvial transport in the Potomac River, and, furthermore, identify this species as a viable candidate organism for hydrophobic organochlorine contaminant biomonitoring in the Chesapeake Bay estuary.

District of Columbia, Maryland, Virginia↗